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布氏田鼠GnRH基因克隆及不同组织和发育阶段的基因表达特征
Molecular Cloning, Tissue Distribution and Expression of GnRH in Different Developmental Stages of Brandt’s vole (Lasiopodomys brandtii)
刘岚,陈燕,李宁,王大伟,任飞,宋英,郭聪,刘晓辉
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作者单位:四川大学生命科学学院中国农业科学院植物保护研究所
中文关键字:布氏田鼠;GnRH;基因克隆;基因表达特征;繁殖调控
英文关键字:Brandt’s vole, GnRH gene, cloning, gene expression, reproductive regulation
中文摘要:促性腺激素释放激素(gonadotrophin releasing hormone,GnRH)由GnRH编码,是调节动物繁殖活动的核心神经内分泌物质。布氏田鼠(Lasiopodomys brandtii)是我国内蒙东部草原区的害鼠之一,具有明显季节性繁殖特征,但其繁殖调控机制仍未完全明确。本研究克隆了来至布氏田鼠下丘脑的GnRH cDNA序列,使用实时荧光定量技术检测了不同组织、不同年龄阶段GnRH mRNA水平。结果表明:克隆获得GnRH cDNA序列497bp,包含开放阅读框(ORF)273bp,编码90个氨基酸和一个终止密码子的GnRH前体。DNA序列比对和氨基酸序列同源性分析表明布氏田鼠下丘脑GnRH基因属于Ⅰ型,与橙腹田鼠GnRH1相似性最高。GnRH mRNA在下丘脑、垂体、睾丸、肾上腺、肠、膀胱,均有表达。雄鼠血清睾酮水平和雌鼠血清雌二醇水平在出生后8周/36周/80周处于高水平,显著高于4周龄鼠,但除出生后80周鼠下丘脑GnRH表达量处于高水平,8周龄/36周龄GnRH表达量与4周鼠均没有显著差异。我们认为可能的原因是,4周龄鼠下丘脑GnRH受GnRH调节剂的中心抑制,而8周龄/36周龄下丘脑GnRH受性类固醇介导的反馈抑制调控,随年龄增加至80周龄,性类固醇介导的反馈抑制消失或者应答时间延长。这些研究为探究布氏田鼠繁殖调控规律提供了更多基础资料。
英文摘要:Gonadotrophin releasing hormone (GnRH) is encoded by GnRH and is the core neuroendocrine substance that regulates the reproductive activity of animals. Brandt’s vole (Lasperodomys brandtii) is one of the rodent species in the grassland of eastern Inner Mongolia, and shows obvious seasonal reproduction, but its reproductive regulation mechanism is still not completely clear. In this study, the GnRH cDNA were cloned from the hypothalamus of Brandt's vole, and the GnRH mRNA of different tissues and different ages were measured using real-time qPCR. The results showed that 497bp of cloned GnRH cDNA sequence contained an open reading frame (ORF) of 273bp, encoding a GnRH precursor of 90 amino acids and a stop codon. Alignment of homologous DNA and amino acid sequences showed that the GnRH of Lasperodomys brandtii belonged to type Ⅰ, and shared the highest identify with Microtus ochrogaste GnRH1. GnRH mRNA was expressed in the hypothalamus, pituitary, testis, adrenal gland, intestine and bladder. The levels of serum testosterone/serum estradiol levels in male/female voles of 8-weeks/36-weeks/80-weeks were significantly higher than those of 4-weeks voles. The expression of GnRH in hypothalamus of 80-weeks voles was in the highest level, while there was no significant difference between 8-weeks/36-weeks voles and 4-weeks voles. We suggested that the possible reason was that the 4-week hypothalamic GnRH was inhibited by the center of the GnRH modulator, while the 8-week/36-week hypothalamic GnRH depended on feedback suppression of steroid. With the increasing age to 80 weeks, sex steroid-mediated feedback suppression declined and response time prolonged, leading to the high GnRH mRNA level in this stage. These results provide basic data for exploring the regulation of reproduction in Brandt’s vole.
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国内统一连续出版物号:51-1193/Q |国际标准出版物号:1000-7083
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